KR101842059B1

KR101842059B1 - Photo-crosslinkable dextran polymer drug delivery with ionic functional group and acrylate group, and method for preparing the same

Info

Publication number: KR101842059B1
Application number: KR1020160014751A
Authority: KR
Inventors: 양수근; 이하늘
Original assignee: 인하대학교 산학협력단
Priority date: 2016-02-05
Filing date: 2016-02-05
Publication date: 2018-03-26
Also published as: KR20170093400A

Abstract

The present invention relates to a photo-crosslinkable dextran polymer drug carrier to which an anionic functional group and an acrylate group are introduced, and a process for preparing the same. The photo-crosslinkable dextran polymer to which the anionic functional group and acrylate group of the present invention are introduced can be easily prepared by introducing a functional group at the same time as the conventional method, and can easily control the composition ratio. The biodegradable dextran Which is biocompatible and can induce an interaction between the anionic functional group and the drug to realize stable drug loading and sustained release, and thus can be used as a drug delivery material containing a drug.

Description

TECHNICAL FIELD The present invention relates to a photo-crosslinkable dextran polymer drug carrier having an anionic functional group and an acrylate group, and a method for preparing the polymer drug carrier,

The present invention relates to a photo-crosslinkable dextran polymer drug carrier to which an anionic functional group and an acrylate group are introduced, and a process for preparing the same.

Dextran is a biocompatible polymer used as a plasma substitute and is a biodegradable polymeric polysaccharide that is degraded in the body by sugar-enzyme. It is a white amorphous solid. It differs in properties depending on the strains it produces, most of which is soluble in water and becomes a white translucent viscous solution, which swells in water and does not dissolve. Partially hydrolyzed is dissolved in water transparently. It does not dissolve in ethanol, ether and so on.

On the other hand, drug delivery systems using ionic interactions have been studied for decades. In 1990, M. Gay Moldenhauer (Moldenhauer MG) reported that anionic ion exchange resins can adsorb theophylline and coat with an ethylcellulose source to control drug release (Journal of Pharmaceutical Sciences, vol. 79, 659-666p) In 1998, Peppas and Nicholas A. used poly (vinyl alcohol) and poly (acrylic acid) as a crosslinking agent for glutaraldehyde and ethylene glycol dimethacrylate It has been described that the drug diffuses and emits by ionic interaction in the bound polymer structure (European Journal of Pharmaceutics and Biopharmaceutics, Vol. 46, 15-29p).

In addition, U.S. Patent No. 2,990,332 discloses a method for producing a sulfonated styrene divinylbenzene copolymerisate in a particle size of 500 μm or less and adsorbing a cationic drug such as amphetamine, atropine, scopolamine and codeine to release a drug from the gastrointestinal tract was technique that can U.S. Patent No. 4,221,778 is an anionic resin Amberlite-XE-69 cation in ^® drug is dextromethorphan (dextromethorphan) by adsorption and be as agarose coated with a echil cellular insolubles continuous drug US Patent No. 4,369,175 discloses an ion exchange drug delivery system prepared by adsorbing vacancies directly from a sulfonic acid resinate having an alkali metal or earth metal salt, Patent Publication No. 4,788,055 discloses that a dextromethorphan plate And a sustained release composition is disclosed. Korean Patent No. 10-0736024 discloses an ion exchange drug delivery system capable of improving the stability of clopidogrel when clopidogrel is adsorbed on a styrene sulfonate polymer or a divinylbenzene styrene sulfonate copolymer. However, the present invention relates to an oral liquid agent for adsorbing a cationic drug such as dextromethorphan or theophylline on a styrene polymer having a sulfonic acid group or by coating with a pharmaceutically applicable polymer or sugar after adsorption to control drug release, There is a problem that it can be applied only to an oral preparation.

In addition, the drug delivery system by ion adsorption is particularly active in the field of microspheres dedicated to liver cancer chemolithography. Jones, C (1989) reported that after ion-adsorbing doxorubicin on ion exchange microspheres, it was effective in the later mouse cancer model (British Journal of Cancer), U.S. Patent No. 7,442,385 discloses that a polyvinyl alcohol polymer is crosslinked And anionic monomers are added to produce anionic polymeric microbeads, it is possible to adsorb a high concentration of a cationic anticancer drug doxorubicin, and when delivered to the hepatic arterial artery, it can simultaneously perform localized chemotherapy with arterial embolization. In addition, patent and technical reports on liver cancer embolization agents and hepatic cancer chemoembolization agents based on polyvinyl alcohol are very diverse, including US Patent Publication No. 7,670,592.

There is collagen coated tris acrylic gelatin micro bead which is currently on the market, but polyvinyl alcohol which is not biodegradable in the body is used as a drug delivery material and there is a disadvantage that the production process is complicated actively, and polyvinyl alcohol When used as a polymer base, it has various problems in terms of biocompatibility and biodegradability. Polyvinyl alcohol microbeads do not biodegrade and remain in the blood vessels and tissues throughout the patient's life. In this case, the microbeads move to the cerebral blood vessels or the pulmonary vessels outside the treatment area and cause serious problems.

Accordingly, the present inventors have made efforts to overcome the problems of the prior art as described above, and as a result, the present inventors have produced a drug delivery system in the form of a microbead in which an anionic functional group and an acrylate group are introduced into a dextran polymer, The preparation method is very simple as compared with the conventional method, and it is easy to arbitrarily control the composition ratio, and there is no concern about the residual polymer by using the biodegradable dextran, and the drug can be stably supported by inducing the interaction with the drug Confirming the fact and completing the present invention.

The present invention provides a drug delivery system comprising an anionic functional group and a photo-crosslinkable dextran polymer introduced with an acrylate group.

The present invention also provides a method for producing the drug delivery system.

The present invention provides a drug carrier comprising an anionic functional group and a photo-crosslinkable dextran polymer to which an acrylate group is introduced.

In addition,

(a) dissolving dextran and an acrylate compound in an organic solvent to prepare dextran acrylate;

(b) dissolving the dextran acrylate and the anionic reagent of step (a) in water to prepare an aqueous solution;

(c) dispersing the aqueous solution of step (b) in an oil phase to prepare an underwater type emulsion; And

(d) exposing the underwater type emulsion of step (c) to UV light to induce photo-crosslinking to prepare a drug delivery vehicle.

The photo-crosslinkable dextran polymer to which the anionic functional group and acrylate group of the present invention are introduced can be easily prepared by introducing a functional group at the same time as the conventional method, and can easily control the composition ratio. The biodegradable dextran Which is biocompatible and can induce an interaction between the anionic functional group and the drug to realize stable drug loading and sustained release, and thus can be used as a drug delivery material containing a drug.

Brief Description of the Drawings Fig. 1 shows NMR spectral results of dextran polymer (A), methacrylic anhydride (B) and dextran methacrylate (C).
2 is a graph showing the results of FT-IR analysis of dextran methacrylate (C) to which 2-acrylamido 2-methylpropanesulfonic acid (A), dextran polymer (B) IR spectrum.
3 is a view showing a method of preparing a drug delivery microbead using a capillary microfluidic system and an optical microscope photograph of a dextran microbead prepared thereby.
FIG. 4A is a diagram showing a result of observing color change after addition of toluidine blue to a microbead having a dextran microbead and an anionic functional group of the present invention. FIG.
FIG. 4B is a view showing a result of observing changes after adding dextran microbeads and a drug (teriparatide) labeled with fluorescence (FITC) to a microbead of text in which an anionic functional group of the present invention is introduced.
FIG. 5 is a graph showing the adsorption rate (A) of teriparatide and the text in which dextran microbeads and the anionic functional groups of the present invention are introduced in the microbeads according to the present invention, (B) of the adsorption amount of paratid.
FIG. 6 is a diagram showing the result of microscopic observation of microbeads before and after adsorption of doxorubicin on the text where anionic functional groups of the present invention are introduced.
FIG. 7 is a graph showing dissolution curves of doxorubicin in microbeads in which the anionic functional group of the present invention is introduced.
FIG. 8 is a graph showing the result of inspecting biocompatibility of a mouse inserted with a microbead into a femoral muscle of a mouse in which an anionic functional group of the present invention is introduced.

The present invention provides a photo-crosslinkable dextran polymer drug carrier to which an anionic functional group and an acrylate group are introduced.

Hereinafter, the present invention will be described in more detail.

The drug delivery system of the present invention can be represented by the following formula (1).

[Chemical Formula 1]

(B) is an anionic functional group introduced for the adsorption of a cationic drug, and the cross-linking of (A) and the cross-linking of (A) B) can be performed simultaneously with light irradiation.

The light-bridging dextran polymer may be in the form of a microbead, and is formed into a microbead by photo-crosslinking.

The diameter of the microbeads may be between 10 μm and 1000 μm, and preferably between 100 μm and 800 μm.

The present invention uses acrylic acid as a photo-crosslinking agent, and the acrylate to be introduced is preferably methyl methacrylate, but not limited thereto. When the photo-crosslinking is induced by using methyl methacrylate, the hydroxyl group (OH-) of dextran is used for the reaction, and the degree of substitution can be controlled according to the molecular weight of dextran.

The molecular weight of the dextran used in the present invention is preferably from 40K to 100K, the substitution ratio of the hydroxy group is preferably from 10 to 50, and more preferably from 25 to 30.

The present invention can introduce an anionic functional group during the process of forming micro beads through photo-crosslinking, thereby omitting a complicated process of synthesizing and purifying the polymer.

The ionic functional group is preferably, but not limited to, 2-acrylamido-2-methylpropane sulfonate.

The weight ratio of the dextran polymer to 2-acrylamido-2-methylpropanesulfonate can be arbitrarily adjusted to prepare various compositions.

The formation of the beads is accomplished through photo-crosslinking of the acrylic acid introduced into dextran and the introduction of the anionic functional groups simultaneously through crosslinking of the acrylic group of the 2-acrylamido-2-methylpropanesulfonate salt with dextran acrylate The acrylic groups not participating in the reaction participate in the photocrosslinking molding of the beads, so that introduction and molding of functional groups are performed in a single step.

The drug may be a cationic drug and may be adsorbed to the drug delivery vehicle by ionic interaction due to the anionic functional group introduced into the drug delivery vehicle of the present invention.

The drug may be an anticancer agent including an anthracycline anticancer agent, an antibiotic agent including tetracycline antibiotics, a protein drug or peptide drug involved in bone formation, more specifically, Doxorubicin hydrochloride, Epirubicin hydrochloride, ), Idarubicin, Daunorubicin, Gemcitabine hydrochloride, Tetracycline hydrochloride, Minocycline hydrochloride, Doxycycline hyclate, Hydrochloric acid Doxycycline hydrochloride, Vancomycin hydrochloride, Teicoplanin, bone morphogenetic protein 2 (BMP-2), bone morphogenetic protein 9 (BMP-9), teriparatide or ex Can be Exenatide

In addition,

(d) inducing photo-crosslinking by exposing the underwater type emulsion of step (c) to UV light to provide an optically crosslinked dextran polymer drug carrier having an anionic functional group and an acrylate group introduced therein do.

The process for preparing the drug delivery system of the present invention will be described in detail as follows.

The step (a) is a step of preparing dextran acrylate by reacting dextran and an acrylate compound. The molecular weight of the dextran is preferably 40 K to 100 K, and the acrylate is preferably methyl methacrylate, Not limited.

In the step (b), the anionic functional group may be 2-acrylamido-2-methylpropanesulfonic acid, but is not limited thereto.

In this case, the aqueous solution may further contain a photoinitiator for photo-crosslinking, and the photoinitiator may include phenyl glyoxylate, acyl phosphine oxide or 2-hydroxyketone, But is not limited thereto.

The step (c) is a step of dispersing the aqueous solution in an oil phase to prepare an underwater type emulsion. The drug delivery system of the present invention can be prepared by various methods, but it can be prepared through the water-in-water type emulsion method .

In the step (d), the drug carrier is prepared by photo-crosslinking. The drug carrier may be in the form of microbeads, and may be formed into microbeads by photo-crosslinking.

The photopolymerizable dextran polymer of the present invention is easily manufactured by introducing a functional group at the same time as the forming method, and is easily biodegradable using biodegradable dextran, and has an anionic functional group and a drug Thereby enabling stable loading and sustained release of the drug.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples.

Example One: Photogenerating Text Acrylate synthesis

Dextran was added to a round flask divided into 40K, 70K and 100K by molecular weight, and 1% w / v LiCl / DMF (N, N-dimethylmethanamide) was added thereto and heated to 80 ° C under reflux to dissolve dextran . Then, the temperature of the dextran solution was lowered to 60 ° C., and triethylamine was added thereto. After 15 minutes, methacrylic anhydride was added thereto, and the reaction was carried out under reflux conditions for 16-20 hours. After the reaction was completed, the reaction mixture was slowly added to cold isopropyl alcohol to precipitate the reaction product, which was then washed 3-4 times with isopropyl alcohol. After washing, the precipitate was separated by centrifugation, dissolved in water and dialyzed again to remove reaction by-products and finally lyophilized to obtain dextran methacrylate. The degree of reaction was controlled by changing the amount of methacrylic anhydride, so that the intensity of the microbeads obtained after photo-crosslinking could be controlled.

NMR spectra of the dextran methacrylate prepared as described above were confirmed, and the results are shown in Fig.

1A to 1C show dextran polymer, methacrylic anhydride and dextran methacrylate, respectively, and their structure was confirmed.

practice Yes 2. Anionic functional groups introduced Photogenerating Text Acrylate Micro Bead Produce

2-1. Manufacture using Encapsulator (B-390, BUCHI)

The mixture of the dextran methacrylate prepared in Example 1, 2-acrylamido 2-methylpropanesulfonic acid and the photoinitiator was dissolved in deionized water, and the mixture was placed in a syringe and passed through a syringe pump at a flow rate of 1-5 ml / min Volumes (1,000-2,500V) and ultrasonic waves (3,000-6,000Hz) were injected into the nozzles while flowing at a rate, and droplets were collected in an oily solution to prepare an underwater type emulsion. In order to maintain the stability of the prepared emulsion, an appropriate amount of an emulsifier (span 60) was added to the oil phase. Then, an optical crosslinking bead was produced by using an encapsulator (B-390, BUCHI) as an anionic text. At this time, MCT oil was used as the oily solution or acetone containing cellulose acetate butyrate (n-butyl acetate) or hydroxypropyl methylcellulose was used. The obtained water-in-oil type emulsion was subjected to light irradiation while magnet agitation to induce hardening to produce final solid microbeads. The solidified beads were washed 2-3 times with an organic solvent (butyl acetate), washed 2-3 times with acetone, and then stored in ethanol.

The FT-IR spectrum of the dextran methacrylate having the 2-acrylamido 2-methylpropanesulfonate thus prepared was confirmed, and the results are shown in FIG.

2A to 2C show dextran methacrylate into which 2-acrylamido 2-methylpropanesulfonic acid, dextran polymer and 2-acrylamido 2-methylpropanesulfonate are introduced, respectively, Respectively.

2-2. Capillary Micro Capillary microfludics Manufacture using

A method of producing acrylate microbeads using textured capillary microfludics is schematically shown in FIG.

As shown in FIG. 3, a microfluidic system was prepared, and a mixture of dextran methacrylate and 2-acrylamido 2-methylpropanesulfonic acid prepared in Example 1 was dissolved in deionized water in an inner glass tube, The aqueous solution was flowed at a flow rate of 1 ml / hour, and MCT oil as a collecting solution was flowed as an external fluid at a flow rate of 30 ml / hour to form a remnant. The formed remnants were collected in MCT oil and crosslinked by light irradiation. A typical photoinitiator was added to the aqueous phase and an appropriate amount of emulsifier (span 60) was added to the emulsion to maintain the stability of the prepared emulsion. The cross-linked cured beads were washed with acetone several times to remove MCT oil and stored in ethanol.

Experimental Example One. Cationic Check for adsorption of drugs

To confirm the adsorption of the cationic drug on the dextran microbeads into which the anionic functional group was introduced in Example 2, a representative cationic indicator drug, toluidine blue, was used.

First, dextran microbeads and toluene blue were added to the microbeads of texts having the anionic functional groups prepared in Example 2, and the color change was observed. The results are shown in FIG. 4A.

As shown in Fig. 4A, it was confirmed that the toluidine blue of the anionic dextran microbead changed from blue to purple. This indicates that the anionic dextran microbeads can adsorb the cationic drug via an electrostatic ionic reaction.

In addition, the dextran microbeads and the text to which the anionic functional group prepared in Example 2 was introduced were observed by adding fluorescence (FITC) labeled drug (teriparatide) to the microbeads, Is shown in Fig. 4B.

As shown in FIG. 4B, in the case of dextran microbeads (B1 and B2), no adsorption of the drug was observed, but in the case of dextran beads (B3 and B4) into which the anionic functional group was introduced, fluorescence- B4).

Experimental Example 2. Confirmation of Adsorption of Cationic Drugs

2-1. Adsorption confirmation of vincycline, BMP2, vancomycin and gemcitabine

Representative cationic drugs were selected and adsorption experiments were performed. First, oxicyclin, BMP2, vancomycin and gemcitabine as cationic drugs were taken in an excessive amount and dissolved in 5 ml of distilled water. Then, the dissolved drug was added to the anionic dextran microbeads (1 ml) prepared in Example 2, and the mixture was gently shaken for 60 minutes at room temperature. Then, the supernatant was taken and the amount of the drug adsorbed on the beads was quantified by ultraviolet absorption analysis The results are shown in Table 1 below.

Cationic drug The amount of drug adsorbed on 1 ml of beads Doxycycline 22 to 7 mg BMP2 5 to 8 ug Vancomycin 38 to 40 mg Gemcitabine 30 to 35 mg

As shown in Table 1, the adsorption of the cationic drug to the anionic dextran microbeads was confirmed.

2-2. Terry Paratide's Adsorption confirmation

Dextran microbeads and the anionic dextran microbeads prepared in Example 2 were added to a dextran photo-crosslinked bead prepared without addition of a cationic peptide drug, teriparatide solution, followed by gentle shaking at room temperature, The adsorption amount was analyzed by HPLC. The results are shown in FIG.

As shown in Fig. 5, it was confirmed that the anionic dextran microbeads adsorbed the cationic drug teriparatide more.

2-3. Doxorubicin Adsorption confirmation

50 mg of doxorubicin was dissolved in 20 ml of distilled water and added to 1 ml of anionic dextran microbeads to conduct adsorption experiments on the drug under the same conditions as Experimental Example 2-1. The amount of adsorbed doxorubicin was 30 ~ Respectively.

The anionic dextran microbeads before and after the adsorption of doxorubicin were observed under a microscope, which is shown in Fig.

As shown in Fig. 6, it was confirmed that doxorubicin was adsorbed on anionic dextran microbeads and turned red.

Experimental Example 3: Elution experiment of cationic drug

The anoxic dextran microbeads adsorbed on doxorubicin in Experimental Example 2-3 were placed in a 500 ml eluate (PBS, pH 7.4) according to Method 2 of US Pharmacopoietic Assay, And the concentration of the drug in the eluate was analyzed by fluorescence analysis. The amount of eluted drug was calculated as cumulative value, and the result is shown in FIG.

As shown in Fig. 7, it was confirmed that doxorubicin eluted continuously for a long time in the anionic dextran microbeads.

Experimental Example 4. Anionic dextran microbeads of Biocompatibility check

The anionic dextran microbeads prepared in Example 2 were injected into the femur muscles of the rats and the injected muscles were cut out two weeks later. The tissue necrosis or inflammatory reaction was observed by H & E staining. The results are shown in Fig. 8 Respectively.

As shown in FIG. 8, it was confirmed that the deposition of inflammatory cells was weak in the vicinity of anionic dextran microbeads, but the biodegradability was considered to be insignificant.

From the above results, it can be seen that the photocrosslinkable dextran microbeads incorporating anionic functional groups and acrylate groups according to the present invention are very simple, biocompatible, and have an interaction with anionic functional groups and drugs And thus it was confirmed that stable drug loading and sustained release could be achieved.

Claims

Dextran polymer; An acrylate group or a methacrylate group; And 2-acrylamido-2-methylpropanesulfonate group,
The dextran polymer is crosslinked to each other through an acrylate group or a methacrylate group,
Wherein the 2-acrylamido-2-methylpropanesulfonate group is chemically bonded to the dextran polymer via an acrylate group or a methacrylate group.

The drug delivery system according to claim 1, wherein the photo-crosslinkable dextran polymer is in a microbead form.

The drug delivery system according to claim 2, wherein the microbead has a diameter of 10 μm to 1000 μm.

delete

The drug delivery system according to claim 1, wherein the drug delivery system is represented by the following formula (1).
[Chemical Formula 1]

The drug delivery system according to claim 1, wherein the drug is a cationic drug.

8. The drug delivery system according to claim 7, wherein the cationic drug is adsorbed on the drug delivery vehicle by ionic interaction.

The pharmaceutical composition according to claim 7, wherein the drug is at least one selected from the group consisting of an anticancer agent comprising an anthracycline-based anticancer agent, an antibiotic including tetracycline antibiotics, a protein drug involved in bone formation, and a peptide drug , Drug delivery system.

10. The method of claim 9, wherein the drug is selected from the group consisting of Doxorubicin hydrochloride, Epirubicin, Idarubicin, Daunorubicin, Gemcitabine hydrochloride, Tetracycline hydrochloride Tetracycline hydrochloride, Minocycline hydrochloride, Doxycycline hyclate, Doxycycline hydrochloride, Vancomycin hydrochloride, Teicoplanin, BMP-2 (bone morphogenetic protein 2 ), BMP-9 (bone morphogenetic protein 9), teriparatide, and exenatide (Exenatide).

(a) dissolving dextran and an acrylate or methacrylate compound in an organic solvent to prepare dextran acrylate;
(b) dissolving dextran acrylate and 2-acrylamido-2-methylpropanesulfonic acid in step (a) in water to prepare an aqueous solution;
(c) dispersing the aqueous solution of step (b) in an oil phase to prepare an underwater type emulsion; And
(d) exposing the underwater type emulsion of step (c) to UV light to induce photocrosslinking.
A method for preparing a drug delivery vehicle comprising the photoconductive dextran polymer of claim 1.

12. The method of claim 11, wherein the drug delivery system in step (d) is in the form of a microbead.

13. The method according to claim 12, wherein the microbead has a diameter of 10 mu m to 1000 mu m.

delete

12. The method according to claim 11, wherein the aqueous solution of step (b) further comprises a photoinitiator.

17. The method of claim 16, wherein the photoinitiator is at least one selected from the group consisting of phenyl glyoxylate, acyl phosphine oxide, and 2-hydroxyketone. Gt; to < / RTI >

12. The method of claim 11, wherein the drug delivery system is represented by the following formula (1).
[Chemical Formula 1]